Dentre os compostos xenobióticos que podem vir a promover efeitos prejudiciais em ecossistemas aquáticos, os fármacos recebem atualmente maior destaque devido à capacidade de persistirem nestes ambientes e também pela escassez de informações de seus efeitos aos diferentes componentes da biota aquática. Dada a presença dessas substâncias, magnitude das concentrações (ng L-1 a μg L-1), e carência de informações sobre seus riscos aos organismos aquáticos nas águas doce no país, o presente estudo estabeleceu como objetivos: i) avaliar a presença dos compostos farmacológicos: diclofenaco de sódio (anti-inflamatório), paracetamol (analgésico) e propranolol (β- bloqueador) nas amostras de água em diferentes pontos selecionados no Reservatório de Guarapiranga-SP; ii) estabelecer o grau de toxicidade aguda destes compostos farmacológicos notoriamente encontrados nas amostras naturais sobre diferentes grupos de organismos aquáticos, como: Ceriodaphnia silvestrii, Daphnia magna, Hydra viridissima e Dugesia tigrina; iii) Avaliar a toxicidade crônica dos fármacos estudados, com a inclusão do antipsicótico clorpromazina, aos cladóceros Ceriodaphnia silvestrii e Daphnia magna; iv) Analisar os efeitos sobre biomarcadores de neurotransmissão, como colinesterases solúveis (ChE); e de estresse e dano oxidativo, tais como catalase (CAT), glutationa-S-transferases (GSTs) e glutationa-peroxidase total e selêniodependente (GPx total e Se-GPx) na espécie D. magna. Os resultados mostraram que as concentrações dos fármacos variaram de 6,04 ng L-1 para o diclofenaco sódico (estação Santa Rita) a 531,4 ng L-1 para o paracetamol (estação Guavirutuba), durante os períodos analisados. Em relação aos resultados obtidos das exposições agudas observou-se uma variabilidade considerável na toxicidade dos dois compostos farmacêuticos estudados, com valores de CE50 e CL50 variando de 0,55 a 123,3 mg L-1, respectivamente, sendo que a espécie D. tigrina foi mais sensível ao fármaco diclofenaco sódico, e ao mesmo tempo apresentou uma maior tolerância ao propranolol. Os dados dos testes de toxicidade crônica mostraram a ocorrência de efeitos adversos na reprodução, mas também efeitos estimulantes para as espécies de Cladocera estudadas. O fármaco propranolol causou um aumento significativo na fecundidade e no parâmetro taxa de crescimento populacional ocorreu um aumento significativo na menor concentração testada para C. silvestrii. Para a espécie D. magna, a clorpromazina e o propranolol causaram uma diminuição significativa na fecundidade e na variável taxa de crescimento populacional. A exposição aos fármacos paracetamol e diclofenaco sódico causou uma inibição na atividade de ChE, Se-GPx e GPx total em D. magna, sendo que o propranolol foi responsável por uma redução significativa nas atividades das duas últimas enzimas mencionadas, e também por um ligeiro aumento na atividade de GSTs. Além disso, somente a atividade da CAT foi alterada de forma significativa na exposição à clorpromazina. Diante do exposto, conclui-se que os fármacos estudados causaram toxicidade aos organismos aquáticos avaliados neste estudo, e que há necessidade de maiores estudos visando integrar a avaliação de risco de fármacos e a proteção dos organismos não-alvo, da ameaça representada pela presença destes produtos no ambiente. / Among the xenobiotic compounds that may cause deleterious effects on aquatic ecosystems, the drugs currently receive greater prominence due to the ability to persist in these environments and also by the lack of information of their effects on different components of the aquatic biota. Given the presence of these, the magnitude of concentrations (ng L-1 to mg L-1), and lack of information about their risks for aquatic organisms in fresh waters in the country, this study has established the following objectives: i) to evaluate the presence of the pharmacutical compounds: diclofenac sodium (anti-inflammatory), paracetamol (analgesic) and propranolol (β-blocker) in water samples at different selected sites in Guarapiranga Reservoir -SP; ii) establish the degree of acute toxicity of these pharmacological compounds notoriously found in natural samples on different groups of aquatic organisms, such as: Ceriodaphnia silvestrii, Daphnia magna, Hydra viridissima and Dugesia tigrina; iii) to assess the chronic toxicity of the drugs studied, with the inclusion of antipsychotic chlorpromazine to cladocerans Ceriodaphnia silvestrii and Daphnia magna; iv) analyze the effects on biomarkers of neurotransmission, such as soluble cholinesterase (ChE); and oxidative stress defense, such as catalase (CAT), glutathione S-transferases (GSTs) and total and selenium-dependent glutathione-peroxidase (total GPx; Se-GPx) in the species D. magna. The results showed that the concentrations of the drugs ranged from 6.04 ng L-1 for diclofenac (Santa Rita station) to 531.4 ng L-1 for paracetamol (Guavirutuba station) during the periods analyzed. Regarding the results obtained from acute exposures a considerable variability in toxicity of the two pharmaceutical compounds studied was observed, and the EC50 and LC50 values ranged from 0.55 to 123.3 mg L-1, and the species D. tigrina was more sensitive to the drug diclofenac, and at the same time had a higher tolerance to propranolol. The data from chronic toxicity tests showed the occurrence of adverse effects on reproduction, but also stimulating effects to the species of Cladocera studied. The drug propranolol caused a significant increase in fertility and regarding the rate of population growth parameter there was a significant increase in the lowest concentration tested for C. silvestrii. For the species D. magna, chlorpromazine and propranolol caused a significant decrease in fertility and the rate of population increase parameter. Exposure to the drugs paracetamol and diclofenac caused an overall inhibition in the activity of ChE, Se-GPx and total GPx in D. magna, and propranolol was responsible for a significant reduction in the activities of the last two enzymes mentioned, and also for a slight increase in the activity of GSTs. Furthermore, only CAT activity was significantly altered in the exposure to chlorpromazine. Given the above, it is concluded that the studied drugs caused toxicity to the aquatic organisms evaluated in this study, which suggests that further studies are necessary to integrate risk assessment of drugs to the protection of non-target organisms of the threat posed by the presence of these products in the environment.

Contaminants may affect marine organisms through various pathways with impacts evident across a variety of spatial and temporal scales. Organisms may encounter short pulsed exposures which contaminate surface waters for hours to days, or more persistent but patchy contamination of benthic habitats throughout their entire life-cycle. This thesis examines the responses of epifauna associated with macroalgae to a pulsed exposure of contaminants (storm-water input) and to chronic contamination via metal accumulation within temperate algal beds. The effects of storm water were monitored during a two-year survey of Sydney Harbour which sampled epifauna before and after heavy rainfall. Epifaunal assemblages declined throughout the harbour following storm events but for the most part these declines were not attributable to storm-water runoff. However, transient (&lt 4 d) and localized impacts of storm water upon physico-chemical characteristics of recipient water and some epifaunal groups were identified around storm drains. A novel field dosing technique tested the relative importance of freshwater and associated metals as causative agents of behavioural avoidance and direct mortality responses. Strong avoidance of storm-water plumes was found which could be entirely explained by freshwater inundation, with no additional effects of metals. No direct mortality was observed following brief exposures. Contaminants introduced by storm water may accumulate within the tissues of macroalgae and potentially pose persistent threats to epifauna. Colonisation of epifauna was reduced on algae with enhanced copper levels, and the nesting behaviour, feeding and survival of an abundant amphipod were all negatively affected by copper load. Subsequent field surveys identified sufficient copper, lead and zinc contamination in Sydney Harbour algal beds to pose direct toxic threats to epifauna. The abundance of herbivorous amphipods correlated negatively with the copper content of a common algal species. However, differences in metal accumulation between algal species resulted in spatially variable levels of contamination. Small-scale patchiness of contaminants within these landscapes may allow populations of mobile species to persist if contaminated hosts are avoided. In summary, epifaunal assemblages appeared resilient to storm-water pulses. Recovery of affected groups was rapid and large fluctuations in abundance appear to be part of the natural flux of epifaunal communities. In contrast, assemblages responded strongly to algal-bound contaminants and this has emerged as an important pathway of contaminant exposure and impact within algal habitats.

Contaminants may affect marine organisms through various pathways with impacts evident across a variety of spatial and temporal scales. Organisms may encounter short pulsed exposures which contaminate surface waters for hours to days, or more persistent but patchy contamination of benthic habitats throughout their entire life-cycle. This thesis examines the responses of epifauna associated with macroalgae to a pulsed exposure of contaminants (storm-water input) and to chronic contamination via metal accumulation within temperate algal beds. The effects of storm water were monitored during a two-year survey of Sydney Harbour which sampled epifauna before and after heavy rainfall. Epifaunal assemblages declined throughout the harbour following storm events but for the most part these declines were not attributable to storm-water runoff. However, transient (&lt 4 d) and localized impacts of storm water upon physico-chemical characteristics of recipient water and some epifaunal groups were identified around storm drains. A novel field dosing technique tested the relative importance of freshwater and associated metals as causative agents of behavioural avoidance and direct mortality responses. Strong avoidance of storm-water plumes was found which could be entirely explained by freshwater inundation, with no additional effects of metals. No direct mortality was observed following brief exposures. Contaminants introduced by storm water may accumulate within the tissues of macroalgae and potentially pose persistent threats to epifauna. Colonisation of epifauna was reduced on algae with enhanced copper levels, and the nesting behaviour, feeding and survival of an abundant amphipod were all negatively affected by copper load. Subsequent field surveys identified sufficient copper, lead and zinc contamination in Sydney Harbour algal beds to pose direct toxic threats to epifauna. The abundance of herbivorous amphipods correlated negatively with the copper content of a common algal species. However, differences in metal accumulation between algal species resulted in spatially variable levels of contamination. Small-scale patchiness of contaminants within these landscapes may allow populations of mobile species to persist if contaminated hosts are avoided. In summary, epifaunal assemblages appeared resilient to storm-water pulses. Recovery of affected groups was rapid and large fluctuations in abundance appear to be part of the natural flux of epifaunal communities. In contrast, assemblages responded strongly to algal-bound contaminants and this has emerged as an important pathway of contaminant exposure and impact within algal habitats.

L’Homme est exposé tout au long de sa vie à de nombreux contaminants présents dans l’environnement et l’alimentation généralement à faibles doses et en mélanges. L’évaluation des risques pose problème dans la mesure où il est bien établi qu’il existe des différences de réponse entre l’homme et l’animal. Quelle que soit la voie d’exposition, de par son rôle majeur dans la biotransformation des xénobiotiques, le foie est considéré comme un organe cible pour de nombreuses classes de produits chimiques potentiellement cytotoxiques, génotoxiques voire cancérogènes. Nous avons utilisé la lignée de cellules hépatiques humaines HepaRG non transformées pour évaluer la toxicité chronique et/ou le pouvoir cancérogène de pesticides et de composés génotoxiques présents dans l’environnement. Cette lignée est la seule connue pour avoir conservé des propriétés proches des hépatocytes humains en culture primaire. Dans une première partie nous avons confirmé le maintien de ses capacités fonctionnelles à confluence par l’analyse du transcriptome et de la biocinétique de 4 médicaments, après traitements quotidiens pendant 14 jours. Nous avons ensuite recherché les effets de mélanges de pesticides après des expositions aiguës et répétées. Nous avons ainsi montré que : 1. l’isomalathion, une impureté majeure du malathion, joue un rôle prépondérant sur la toxicité hépatique de ce dernier et qu’il inhibe la carboxylesterase impliquée dans le métabolisme des deux composés; 2. l’endosulfan et le méthoxychlore, deux organochlorés métabolisés par les CYP3A4 et 2B6, agissent de manière synergique sur leur cytotoxicité après une exposition unique ou répétée. De plus, alors que l'activité du CYP3A4 est inhibée de manière réversible par l’endosulfan, le méthoxychlore l’augmente. En revanche, l’activité du CYP2B6 est induite par les deux pesticides. Lorsqu’ils sont en mélange équimolaire un effet additif ou antagoniste est observé sur l'activité du CYP3A4 et du CYP2B6 respectivement quelle que soit la durée de l’exposition. Enfin, dans une troisième partie, nous avons exposé des cellules HepaRG pendant une quinzaine de passages à de faibles doses de deux contaminants génotoxiques nécessitant une bioactivation, l’aflatoxine B1 et une amine aromatique hétérocyclique, le PhiP, et démontré l’acquisition de propriétés de cellules transformées (par exemple croissance sur agar, migration dans le test de griffure et surexpression de gènes associés au cancer). Au total, nos résultats démontrent tout l’intérêt que représente la lignée hépatique humaine HepaRG métaboliquement compétentes pour l’étude de la toxicité chronique et/ou le potentiel cancérogène des contaminants de l’environnement. Ils ont permis de mettre en évidence d’interactions entre des pesticides en mélanges binaires et pour la première fois d’analyser le potentiel cancérogène de contaminants génotoxiques dans une lignée hépatique humaine. / Humans are exposed throughout their life to many environmental and food contaminants, usually at low doses and in mixtures. Risk assessment remains questionable as it is well established that there are differences in the response to chemicals between humans and animals. Regardless of the route of exposure, due to its major role in xenobiotic biotransformation, the liver is considered as a target organ for many classes of chemicals potentially cytotoxic, genotoxic or carcinogenic. We used the HepaRG cell line to evaluate chronic toxicity and/or carcinogenicity of pesticides and genotoxic compounds. This cell line is the only one known to exhibit properties similar to those of human hepatocytes in primary culture. In the first part we confirmed the maintenance of functional capacities of these cells at confluence by transcriptomic and biokinetic analysis of several drugs after a 14-day treatment. We then investigated the effects of mixtures of pesticides after acute and repeated exposures. We showed that : 1. Isomalathion, a major impurity of malathion, played a leading role on liver toxicity and inhibited carboxylesterase that is involved in the metabolism of these two compounds; 2. Endosulfan and methoxychlor, two organochlorines, metabolized by CYP3A4 and CYP2B6, acted synergistically on their cytotoxicity after single or repeated exposure. Moreover, whereas activity of CYP3A4 was reversibly inhibited by endosulfan and increased by methoxychlor. By contrast, CYP2B6 activity was induced by these two pesticides while in equimolar mixtures, they caused additive or antagonistic effects on CYP3A4 and CYP2B6 activities respectively, regardless of the duration of exposure. Finally, in the third part, we exposed HepaRG cells for up to 15 passages to low doses of two genotoxic contaminants which required bioactivation, aflatoxin B1 and heterocyclic aromatic amine, PhIP, and demonstrated the appearance of properties of transformed cells (e.g. growth on agar, cell migration in the wound healing test and overexpression of a number of genes associated with cancer). Altogether, our results demonstrate the great potential interest that represents the metabolically competent human liver cell line HepaRG for the study of chronic toxicity and/or carcinogenic potential of environmental contaminants. They highlight possible interactions between pesticides in binary mixtures and for the first time, demonstrate that the carcinogenic potential of genotoxic contaminants can be analyzed in an human hepatic cell line.

Muitas substâncias descartadas no meio ambiente não são totalmente degradadas, podendo assim persistir no ambiente. Diversos compostos são continuamente introduzidos no ambiente podendo afetar a biota e inclusive o homem. Os fármacos são alguns desses compostos que depois de descartados podem chegar nos corpos de águas naturais, e dentre eles merecem especial atenção os hormônios sintéticos utilizados em larga escala por mulheres em todo o mundo, na forma de contraceptivos orais. O hormônio sintético 17α-etinilestradiol é um micropoluente no ambiente aquático, que pode causar distúrbios na reprodução de diversos organismos atuando como um desregulador endócrino. O presente estudo teve como objetivo principal analisar o efeito do hormônio sintético 17α-etinilestradiol sobre o cladócero Daphnia magna, por meio de testes ecotoxicológicos. Testes de toxicidade crônica foram realizados em duas gerações consecutivas deste microcrustáceo (F0 e F1). Para os testes utilizaram-se neonatas com menos de 24 horas de idade, 6 concentrações do hormônio e dois controles. Foram estabelecidas 10 réplicas com 1 indivíduo por réplica. O ensaio foi realizado em incubadora com temperatura de 25 ± 1°C e fotoperíodo de 12h claro:12h escuro, com duração de 11 (F0) e 13 dias (F1), com término coincidindo com o nascimento das neonatas da terceira ninhada no controle. Os resultados evidenciaram que a exposição ao hormônio diminuiu a fecundidade de Daphnia magna nas quatro maiores concentrações de etinilestradiol na F0 e na concentração de 1000 μg L-1 da F1, revelando maior resistência ao contaminante na segunda geração. Na maior concentração do composto, o tempo para a produção das duas primeiras ninhadas foi maior na geração F1, quando comparada ao controle. Na concentração de 250 μg L-1 verificou-se a ocorrência de um indivíduo intersexo, apresentando tanto características de macho como de fêmea. Os resultados deste estudo evidenciaram que o 17α-etinilestradiol afeta a reprodução de Daphnia magna, e que também pode afetar a reprodução de diferentes invertebrados aquáticos, o que, a longo prazo pode causar danos às populações e comunidades aquáticas, diminuindo as populações e podendo até extingui-las eventualmente. / Many substances are discarded in the environment and not completely degraded, thus persisting in the environment. Some of these are continuously introduced in the environment, affecting the biota, including man. Pharmaceutical drugs are some of these compounds that after discarded can occur in natural water bodies and among them the synthetic hormones deserve special attention for being used in large scale by women world widely, as oral contraceptives. The synthetic hormone 17α-ethinyl estradiol is therefore a micropoluent in the aquatic environment, i. e. found in low concentrations that can cause deleterious effects in the reproduction of many organisms, acting as an endocrine disruptor. The present study had as main objective to analyze the effect of the synthetic estrogen 17α-ethinyl estradiol on the cladoceran Daphnia magna, by carrying out ecotoxicological tests. Chronic toxicity tests were performed on two consecutive generations of this microcrustacean (F0 and F1). In order to perform the tests, neonates aged less than 24 hours, 6 hormones concentrations and two types of controls were used. Ten replicates were established with one individual each. The test was performed in a growth chamber at the constant temperature of 25 ± 1°C and 12 h light:12 h dark photoperiod, had the duration of 11 and 13 days for the F1 and F0 generations, respectively, coinciding with the birth of the third brood in the control. The results evidenced that the exposition to the hormone decreased D. magna fecundity in the four highest of ethinyl estradiol in F0, and in the concentration 1000 μg L-1 for the F1, indicating resistance increase in the second generation. In the highest concentration of this compound the time for the production of the first two broods were higher in the F1 generation as compared with the controls. In the hormone concentration of 250 μg L-1 the occurrence of an intersex individual was verified, simultaneously presenting characteristics of male and female. The results of this study evidenced the 17α-ethinyl estradiol affect the reproduction of Daphnia magna, and can affect the reproduction of other aquatic invertebrates that at long term can cause damages to aquatic populations and communities by diminishing populations and eventually leading them to the extinction.

Made available in DSpace on 2016-01-26T18:55:44Z (GMT). No. of bitstreams: 1 Mozart Alves Goncalves Filho.pdf: 823168 bytes, checksum: 9a480c850a219c880f8d278e6850d549 (MD5) Previous issue date: 2015-06-26 / Cancer is a universal disease, with high mortality rates in some cases. It is the second leading cause of death in the Western world, and go to the first place around the year 2020. Cancer has genetic and environmental causes, one of them the ingestion of heavy metals such as cadmium. There are few studies evaluating the gastrointestinal toxicity of cadmium. There is no consensus in the literature on the treatment of cadmium toxicity. We need simple methods to avoid its effects. Objective: The objective of this study was to evaluate the lesions caused by cadmium poisoning in the digestive tract and the possible effect of the drinking water pH in the initiation of these lesions. Methods: For this study, 90 male Wistar rats were used, divided into 6 groups: A - 15 rats that received 400 mg / L cadmium chloride (CdCl2) in drinking water at a neutral pH of 7.0; B - 15 rats that received CdCl2 (400 mg / L) in drinking water at an acidic pH of 5.0; C - 15 rats that received CdCl2 (400 mg / L) in drinking water at a basic pH of 8.0; D - 15 rats that received water at an acidic pH of 5.0; E - 15 rats that received water at a basic pH of 8.0; and F - 15 rats that received water at a neutral pH of 7.0. All animals were euthanized after 6 months. Fragments of the esophagus, stomach, small intestine and large intestine of each rat were removed for microscopic analysis. Results: There were microscopic changes neither in the esophagus nor in the small and large intestine. Only cadmium exposed animals showed mild dysplasia of the gastric mucosa (p= 0.012), regardless of the pH (p> 0.05). Conclusion: The cadmium led to the formation of dysplastic lesions in the gastric glandular epithelium, regardless of water pH. / Câncer é uma doença universal, com taxa de mortalidade elevada em alguns casos. É a segunda causa de morte no mundo ocidental, e passará para primeiro lugar por volta do ano de 2020. Tem causas genéticas e ambientais, sendo uma delas a ingestão de metal pesado, como cádmio. Há poucos estudos avaliando a toxicidade gastrointestinal do cádmio. Não há consenso na literatura sobre o tratamento da toxicidade por cádmio. Há necessidade de métodos simples de evitar seus efeitos. Objetivo: O objetivo deste estudo foi avaliar as lesões causadas por intoxicação por cádmio no aparelho digestório e os possíveis efeitos do pH da água na gênese destas lesões. Material e métodos: Para este estudo, foram utilizados 90 ratos Wistar, machos. Os animais foram distribuídos em 6 grupos (n=15): A solução de cloreto de cádmio (400mg/L) na água de beber com pH neutro (pH 7,0); B solução de cloreto de cádmio (400mg/L) na água de beber com pH ácido (pH 5,0); C solução de cloreto de cádmio (400mg/L) na água com pH básico (pH 8,0). D água de beber com pH ácido (pH 5,0); E água de beber com pH básico (pH 8,0); F água com pH neutro (pH 7,0). Animais de todos os grupos foram eutanasiados após 6 meses. Foram retirados fragmentos do esôfago, estômago, intestino delgado e intestino grosso de cada rato para análise microscópica. Resultados: Não foram observadas alterações microscópicas no esôfago nem no intestino delgado e grosso. Somente animais expostos ao cádmio apresentaram displasia leve da mucosa gástrica (p= 0,012), porém sem diferença com relação aos diferentes pH da água (p>0,05). Conclusão: O cádmio levou a formação de lesões displásicas no epitélio glandular gástrico, independente do pH da água.

Made available in DSpace on 2016-07-18T17:53:15Z (GMT). No. of bitstreams: 1 Mozart Alves Goncalves Filho.pdf: 823168 bytes, checksum: 9a480c850a219c880f8d278e6850d549 (MD5) Previous issue date: 2015-06-26 / Cancer is a universal disease, with high mortality rates in some cases. It is the second leading cause of death in the Western world, and go to the first place around the year 2020. Cancer has genetic and environmental causes, one of them the ingestion of heavy metals such as cadmium. There are few studies evaluating the gastrointestinal toxicity of cadmium. There is no consensus in the literature on the treatment of cadmium toxicity. We need simple methods to avoid its effects. Objective: The objective of this study was to evaluate the lesions caused by cadmium poisoning in the digestive tract and the possible effect of the drinking water pH in the initiation of these lesions. Methods: For this study, 90 male Wistar rats were used, divided into 6 groups: A - 15 rats that received 400 mg / L cadmium chloride (CdCl2) in drinking water at a neutral pH of 7.0; B - 15 rats that received CdCl2 (400 mg / L) in drinking water at an acidic pH of 5.0; C - 15 rats that received CdCl2 (400 mg / L) in drinking water at a basic pH of 8.0; D - 15 rats that received water at an acidic pH of 5.0; E - 15 rats that received water at a basic pH of 8.0; and F - 15 rats that received water at a neutral pH of 7.0. All animals were euthanized after 6 months. Fragments of the esophagus, stomach, small intestine and large intestine of each rat were removed for microscopic analysis. Results: There were microscopic changes neither in the esophagus nor in the small and large intestine. Only cadmium exposed animals showed mild dysplasia of the gastric mucosa (p= 0.012), regardless of the pH (p> 0.05). Conclusion: The cadmium led to the formation of dysplastic lesions in the gastric glandular epithelium, regardless of water pH. / Câncer é uma doença universal, com taxa de mortalidade elevada em alguns casos. É a segunda causa de morte no mundo ocidental, e passará para primeiro lugar por volta do ano de 2020. Tem causas genéticas e ambientais, sendo uma delas a ingestão de metal pesado, como cádmio. Há poucos estudos avaliando a toxicidade gastrointestinal do cádmio. Não há consenso na literatura sobre o tratamento da toxicidade por cádmio. Há necessidade de métodos simples de evitar seus efeitos. Objetivo: O objetivo deste estudo foi avaliar as lesões causadas por intoxicação por cádmio no aparelho digestório e os possíveis efeitos do pH da água na gênese destas lesões. Material e métodos: Para este estudo, foram utilizados 90 ratos Wistar, machos. Os animais foram distribuídos em 6 grupos (n=15): A solução de cloreto de cádmio (400mg/L) na água de beber com pH neutro (pH 7,0); B solução de cloreto de cádmio (400mg/L) na água de beber com pH ácido (pH 5,0); C solução de cloreto de cádmio (400mg/L) na água com pH básico (pH 8,0). D água de beber com pH ácido (pH 5,0); E água de beber com pH básico (pH 8,0); F água com pH neutro (pH 7,0). Animais de todos os grupos foram eutanasiados após 6 meses. Foram retirados fragmentos do esôfago, estômago, intestino delgado e intestino grosso de cada rato para análise microscópica. Resultados: Não foram observadas alterações microscópicas no esôfago nem no intestino delgado e grosso. Somente animais expostos ao cádmio apresentaram displasia leve da mucosa gástrica (p= 0,012), porém sem diferença com relação aos diferentes pH da água (p>0,05). Conclusão: O cádmio levou a formação de lesões displásicas no epitélio glandular gástrico, independente do pH da água.

The problem of high proportion of zeroes has long been an interest in data analysis and modeling, however, there are no unique solutions to this problem. The solution to the individual problem really depends on its particular situation and the design of the experiment. For example, different biological, chemical, or physical processes may follow different distributions and behave differently. Different mechanisms may generate the zeroes and require different modeling approaches. So it would be quite impossible and inflexible to come up with a unique or a general solution. In this dissertation, I focus on cases where zeroes are produced by mechanisms that create distinct sub-populations of zeroes. The dissertation is motivated from problems of chronic toxicity testing which has a data set that contains a high proportion of zeroes. The analysis of chronic test data is complicated because there are two different sources of zeroes: mortality and non-reproduction in the data. So researchers have to separate zeroes from mortality and fecundity. The use of mixture model approach which combines the two mechanisms to model the data here is appropriate because it can incorporate the mortality kind of extra zeroes. A zero inflated Poisson (ZIP) model is used for modeling the fecundity in <i> Ceriodaphnia dubia</i> toxicity test. A generalized estimating equation (GEE) based ZIP model is developed to handle longitudinal data with zeroes due to mortality. A joint estimate of inhibition concentration (ICx) is also developed as potency estimation based on the mixture model approach. It is found that the ZIP model would perform better than the regular Poisson model if the mortality is high. This kind of toxicity testing also involves longitudinal data where the same subject is measured for a period of seven days. The GEE model allows the flexibility to incorporate the extra zeroes and a correlation structure among the repeated measures. The problem of zero-heavy data also exists in environmental studies in which the growth or reproduction rates of multi-species are measured. This gives rise to multivariate data. Since the inter-relationships between different species are imbedded in the correlation structure, the study of the information in the correlation of the variables, which is often accessed through principal component analysis, is one of the major interests in multi-variate data. In the case where mortality influences the variables of interests, but mortality is not the subject of interests, the use of the mixture approach can be applied to recover the information of the correlation structure. In order to investigate the effect of zeroes on multi-variate data, simulation studies on principal component analysis are performed. A method that recovers the information of the correlation structure is also presented. / Ph. D.

Darbo tikslas Aprašyti profesinių ligų tendencijas Lietuvoje 1973-1978 m. Uždaviniai 1. Nustatyti profesinių ligų skaičiaus ir sergamumo rodiklio pokyčius 1973-1978 m. Lietuvoje. 2. Nustatyti profesinių ligų pasiskirstymą Lietuvoje 1973-1978 m. pagal rizikos veiksnius, ligų grupes, ekonominės veiklos rūšis bei apskritis. 3. Nustatyti profesinių ligų pasiskirstymą Lietuvoje 1973-1978 m. pagal lytį ir darbo stažą. Metodika Duomenys apie 1303 profesinių ligų atvejus buvo surinkti iš 6 LCVA bylų, kuriose yra surinktos 1973-1978 m. tuometinės LTSR SAM ketvirtinės profesinių ligų ataskaitos bei teritorinių SES pranešimai apie profesines ligas. Aprašomoji statistika atlikta Microsoft Excel 2003, Microsoft Excel 2003, WinPepi 11.18, SPSS 17.0 programomis, naudojant &#967;2 ir tikslųjį Fišerio kriterijų. Išvados apie santykinio požymių skirtumo statistinį reikšmingumą buvo daromos, kai p &#8804; 0,05. Rezultatai ir išvados Vidutinis sergamumo profesinėmis ligomis Lietuvoje 1973-1978 m. rodiklis buvo 16,3 / 100 tūkst. dirbančiųjų. Daugiausiai profesinių ligų buvo įtakotos fizikinių veiksnių (51,04 proc.). Daugiausiai registruota ausies (35,5 proc.) ir jungiamojo audinio ir skeleto raumenų sistemos (17,2 proc.) ligų. Daugiausiai profesinių ligų registruota Kauno (31,2 proc.) apskrityje bei apdirbamosios gamybos darbuotojams (64,1 proc.). Vyrams daugiau buvo registruota ūmių profesinių ligų, o moterims – lėtinių. Moterys daugiau sirgo ausies bei nervų sistemos ligomis, o vyrai jungiamojo... [toliau žr. visą tekstą] / Purpose of Study Describe tendencies of occupational diseases in Lithuania in 1973-1978. Objectives 1. Set the number of occupational diseases and the incidence rate of change in Lithuania in 1973-1978. 2. Determine the distribution of occupational diseases according to risk factors, disease groups, industry and region in Lithuania in 1973-1978. 3. Determine the distribution of occupational diseases by sex and total experience of working in Lithuania in 1973-1978. Methodology Data on occupational diseases about 1303 cases were collected from LCVA of 6 files, in which Lithuanian SSR Ministry of Health quarterly reports and territorial Sanitary Epidemiological Station reports about occupational diseases are archived. Descriptive statistics performed in Microsoft Excel 2003, Microsoft Excel 2007, WinPepi 11.18, SPSS 17.0 programs using &#967;2 and Fisher's exact test. Conclusions about the relative signs of the difference between statistical significance were made at p &#8804; 0.05. Conclusions and Results The average incidence rate of occupational diseases in Lithuania in 1973-1978 was 16.3 / 100 000 workers. Most occupational diseases have been influenced by physical factors (51.04%). Majority of them was ear (35.5%) and connective tissue and musculoskeletal system (17.2%) diseases. Most occupational diseases registered in Kaunas (31.2%) and for manufacturing workers (64.1%). There have been registered more acute occupational disease for men, and chronic for women. Women had... [to full text]

Introdução: A quimiorradioterapia (QRT) concomitante baseada em cisplatina é uma opção de tratamento empregada para os pacientes com carcinoma epidermoide de cabeça e pescoço (CECCP) localmente avançado e com bom performance status, seja em caráter adjuvante ou definitivo. O ganho de sobrevida com esta modalidade de tratamento é acompanhado de aumento das toxicidades agudas em comparação com a radioterapia isolada. A ocorrência de toxicidades tardias é menos reportada na literatura e incluem xerostomia, disfagia, hipotireoidismo, ototoxicidade, fístula/necrose cutânea, dentre outras. Tais sequelas tardias podem comprometer a qualidade de vida do sobrevivente ao CECCP. Objetivos: Verificar a prevalência de toxicidades tardias em sobreviventes ao CECCP tratados com QRT baseada em cisplatina. Métodos: Estudo transversal, uni-institucional, que incluiu de forma sequencial pacientes acima de 18 anos, tratados para CECCP (sítios primários: nasofaringe, orofaringe, cavidade oral, hipofaringe e laringe) e que haviam recebido QRT adjuvante ou definitiva, baseada em cisplatina. Estes pacientes estavam em seguimento há pelo menos 2 anos, sem evidência de doença. Os pacientes realizaram audiometria, endoscopia digestiva alta (EDA), nasofibrolaringoscopia da deglutição (NFL), exames laboratoriais (toxicidade tireoidiana e renal). Os pacientes incluídos também foram examinados clinicamente e as toxicidades apresentadas foram graduadas de acordo com a escala de toxicidades tardias do RTOG/EORTC. Os sobreviventes foram ainda avaliados quanto à percepção das toxicidades através de um inventário de sintomas e responderam questionários de qualidade de vida. Resultados: De janeiro de 2014 a fevereiro de 2017, 120 pacientes assinaram o TCLE. A idade mediana dos pacientes é 59 anos (21-78), com predomínio do sexo masculino (73%) e da cor branca (58%). Antecedente de tabagismo foi referido por 80% da amostra e de etilismo por 63%. Referente ao sítio primário, a maioria dos pacientes apresenta tumor em orofaringe (42%), seguido por laringe (23%) e cavidade oral (19%). O tempo de seguimento mediano é 42 meses (24-125). Há predomínio de pacientes com doença localmente avançada, tumores T3/T4 em 75% da amostra e N+ em 72%. A dose mediana de cisplatina recebida durante a concomitância foi 300 mg/m² (100-300) e de radioterapia foi 70 Gy (60-70,4). A QRT foi oferecida em caráter adjuvante em 49% da amostra. As toxicidades mais relatadas pelos pacientes foram: xerostomia (83%), alteração na voz (74%), saliva pegajosa (73%) e disfagia (73%). Ao se graduar as toxicidades conforme escala do RTOG/EORTC, verificou-se que a maioria das toxicidades apresentadas eram de graus leves, 1 ou 2. EDA encontrou estenose faríngea em 10% dos pacientes e NFL identificou fibrose em 37% dos sobreviventes. Dos pacientes submetidos a audiometria, 42% apresentaram perda auditiva de possível causa ototóxica. Cerca de 14% dos sobreviventes apresentam clearance de creatinina estimado < 60 mL/min/1,73m². Conclusões: Toxicidades tardias foram frequentemente reportadas pelos sobreviventes ao CECCP após QRT, porém, na maioria das vezes, de intensidade leve (graus 1 ou 2). Após a QRT, um seguimento cuidadoso é essencial para diagnóstico precoce e reabilitação a essas toxicidades, a fim de preservar a funcionalidade e qualidade de vida dos pacientes / Background: Cisplatin based CRT is the standard therapy for patients with locally advanced HNSCC with good performance status either as adjuvant or as definitive treatment. The survival gain with this treatment modality is accompanied by an increase in acute toxicities in comparison with isolated radiotherapy. The occurrence of LT is less reported in the literature and includes xerostomia, dysphagia, hypothyroidism, ototoxicity, cutaneous fistula / necrosis, among others. Such late sequelae may compromise the survivor\'s quality of life. Endpoints: To verify the prevalence of late toxicities in HNSCC survivors treated with cisplatin based CRT. Methods: A cross-sectional study that sequentially included patients over 18 years of age who were previously treated for HNSCC (primary sites: nasopharynx, oropharynx, oral cavity, hypopharynx and larynx) and who had received either adjuvant or definitive cisplatin based CRT. These patients were in follow-up for at least 2 years, with no evidence of disease. The patients underwent audiometry, upper GI endoscopy, nasopharyngolaryngoscopy (NPL), laboratory tests (thyroid and kidney toxicity). The included patients were also clinically assessed for mucous membrane, skin, subcutaneous tissue, salivary gland, larynx and esophagus LT according to the RTOG/EORTC Late Radiation Morbidity Scoring Schema. All patients answered a questionnaire about their perception of LT through a symptoms inventory and also answered QoL questionnaires. Results: From January 2014 to February 2017, 120 patients signed the informed consent form. The mean age of the patients is 59 years (21-78), predominantly male (73%) and white (58%). Previous smoking habits were reported by 80% of the sample and alcohol consumption by 63%. Most common primary sites were oropharynx (42%), followed by larynx (23%) and oral cavity (19%). The median follow-up time is 42 months (24-125). There was a predominance of locally advanced disease, T3 / T4 tumors in 75% of the sample and N + in 72%. The median cisplatin dose during concomitance was 300 mg/m² (100-300) and the median radiotherapy delivered dose was 70Gy (60-70.4). CRT was delivered as an adjuvant treatment in 49% of the sample. The most frequently selfreported LT were xerostomia (83%), voice disorders (74%), sticky saliva (73%) and dysphagia (73%). Assessing the toxicities according to the RTOG / EORTC scale most of them were mild, grade 1-2. Upper GI endoscopy diagnosed stenosis in 10% of the patients and NPL identified fibrosis in 37% of the survivors. Audiometry identified ototoxic hearing loss in 42% of the sample. About 14% of the survivors present chronic kidney disease (an estimated creatinine clearance < 60 mL/min/1.73m²). Conclusion: High rates of self-reported LT were detected although most of them seem to be mild. After CRT, a close follow-up of HNSCC patients is essential for early diagnosis, treatment of these late sequelae and rehabilitation, in order to preserve QoL and functionality and to avoid lifethreatening conditions and social reclusion